LOCUS       sequence-296063-        5199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry-tagged HRS 2xFYVE in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5199)
  AUTHORS   Sneeggen M, Pedersen NM, Campsteijn C, Haugsten EM, Stenmark H, 
            Schink KO
  TITLE     WDFY2 restrains matrix metalloproteinase secretion and cell invasion
            by controlling VAMP3-dependent recycling.
  JOURNAL   Nat Commun. 2019 Jun 28;10(1):2850. doi: 10.1038/s41467-019-10794-w.
  PUBMED    31253801
REFERENCE   2  (bases 1 to 5199)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-06-28"; volume: "10(1):2850. doi"; pages: " 
            10.1038/s41467-019-10794-w"
COMMENT     Sequence Label: pmCherry-2xFYVE
FEATURES             Location/Qualifiers
     source          1..5199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             612..1319
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(757..775)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1025..1044
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1986..2107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2023..2042)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2077..2096
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2114..2569)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2251..2272)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2463..2482
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2596..2700
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2702..3059
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2702..2893
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2725..2745)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2910..3045
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3094..3888
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3094..3888
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3148..3167)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3758..3777
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4076..4095)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4120..4167
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4496..5084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4985..5004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      601 cggtcgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
      661 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
      721 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
      781 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
      841 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
      901 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
      961 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     1021 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     1081 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     1141 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1201 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     1261 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     1321 actcagatct cgagctcaag cttcgaattg aattcgaaag tgatgccatg ttcgctgctg
     1381 aaagagcccc tgactgggtg gatgctgagg aatgccatcg gtgcagagta cagtttgggg
     1441 tggtgacccg caagcatcac tgccgagcat gtgggcagat cttctgtggc aagtgctcct
     1501 ccaagtactc caccatcccc aagttcggca ttgagaagga ggtgcgcgtg tgtgagccct
     1561 gctatgagca gctgaacaag aaggcacaag gacaaggatc cgaaagtgat gccatgttcg
     1621 ctgctgaaag agcccctgac tgggtggatg ctgaggaatg ccatcggtgc agagtacagt
     1681 ttggggtggt gacccgcaag catcactgcc gagcatgtgg gcagatcttc tgtggcaagt
     1741 gctcctccaa gtactccacc atccccaagt tcggcattga gaaggaggtg cgcgtgtgtg
     1801 agccctgcta tgagcagctg aacaagaagg cataagtcga cggtaccgcg ggcccgggat
     1861 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     1921 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     1981 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2041 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2101 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2161 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2221 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2281 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2341 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2401 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2461 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2521 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     2581 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2641 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2701 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2761 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2821 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2881 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2941 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3001 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     3061 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3121 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3181 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3241 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     3301 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3361 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3421 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3481 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3541 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3601 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     3661 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3721 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3781 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3841 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     3901 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     3961 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4021 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     4081 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4141 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4201 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     4261 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     4321 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     4381 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4621 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5101 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5161 cccctgattc tgtggataac cgtattaccg ccatgcatt
//
